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Biomedical subjects

T Kishida

Publications and source records attributed to T Kishida.

At least 109 records · Page 6Linked to original sources
PubMed1996

Effect of the oral administration of Lactobacillus brevis subsp. coagulans on interferon-alpha producing capacity in humans.

OBJECTIVE: This study was designed to determine whether the oral administration of Lactobacillus brevis subsp. coagulans modulates immunological responses in human subjects, and whether there are differences in response between live and heat-treated preparations. METHODS: The effect of the oral administration of Lactobacillus brevis subsp. coagulans (Labre) for 4 weeks on the interferon-alpha (IFN-alpha) producing capacity of apparently healthy subjects was evaluated. Sixty volunteers were divided into five groups for the determination of virus-induced IFN-alpha production in response to various doses of live and to heat-killed Labre. 2-5A synthetase activity was measured to detect trace amounts of IFN production. Routine blood tests were also performed to determine the state of health of the subjects involved in this study and to test any side effects of Labre treatment. RESULTS: The oral administration of live Labre showed a statistically significant increase in IFN-alpha production at 2 weeks (p < 0.05) and at 4 weeks (p < 0.05) in the group receiving 600 million bacteria/day and at 4 weeks (p < 0.05) in the group receiving 300 million bacteria/day. In particular, IFN-alpha production in those with initially low levels rose significantly when either 300 million or 600 million bacteria/day were ingested. Consumption of heat-killed Labre 300 million bacteria/day did not result in a statistically significant change in IFN-alpha production. The level of 2-5A synthetase activity remained the same in the control and experimental groups. CONCLUSIONS: Oral administration of live Labre significantly increased IFN-alpha production in a dose-dependent manner. Labre intake tended to be most beneficial in subjects with initially low levels of IFN-alpha production. Heat-treated Labre did not elicit a response similar to that of the live bacteria.

2',5'-Oligoadenylate Synthetase↗
PubMed1996

Determination of interferon-alpha-producing capacity in whole blood cultures from patients with various diseases and from healthy persons.

To assess the clinical value of determination of the interferon (IFN)-producing capacity of patients, IFN production induced by Sendai virus (HVJ) in vitro was measured in cell cultures of whole blood from patients with various diseases. IFN production in patients with lung cancer, myelodysplastic syndromes, noninsulin-dependent diabetes mellitus, pulmonary tuberculosis, and asymptomatic HIV-1 infection was lower than that in healthy persons. Furthermore, periodic measurements of IFN production revealed decreasing IFN producing capacities in patients with lung cancer with progression of the tumor stage. However, increased IFN-producing capacities were observed in patients with tuberculosis after standard therapy. Further experiments showed that the main type of IFN induced in whole blood cultures was IFN-alpha, and decreased IFN production in patients did not result from a decreased number of leukocytes but rather from an impairment of cellular IFN production. The evaluation of IFN production in whole blood cell cultures may be a feasible method of assessing the impaired immune status.

Adult↗
PubMed1996

Parturition in six renal allograft recipients.

Between 1983 and 1994, we studied renal function and neonatal conditions for eight pregnancies and births to six women who had received renal transplants in order to assess the effect of an allograft on pregnancy and its outcome. The gestation period was 34 to 39 weeks (mean 36 weeks and 4 days), and four pregnancies ended before term. All eight babies were delivered by cesarean section. Intrauterine growth retardation (IUGR) was found in both babies of one woman who had been treated with conventional (without cyclosporin) immunosuppression. The serum creatinine level did not change during gestation in any of the women but was elevated after delivery in four. Four mothers suffered from proteinuria (25-364 mg/dl) during gestation, but the proteinuria disappeared after delivery in all but one case. The one exception, persistent proteinuria of 100-200 mg/dl, was assumed to result from the recurrence of the original renal disease (lgA nephropathy). The reduction of creatinine clearance and hydronephrosis of one graft noted during gestation were later reversed. None of the eight babies (four females and four males) was congenitally malformed, and their Apar scores were 6 to 9 (median 8). They are now 3 months to 11 years old, and seven of them are healthy and show good growth. One of the two IUGR babies has not grown well; her weight and height are more than 1 SD below the mean for her age, and she is mentally retarded and suffers from muscle weakness. Compared with dialysis patients, female renal allograft recipient have a better quality of life because they can safely deliver a child if they observe the criteria for pregnancy established for renal allogaft recipients.

Child↗
PubMed1996

Cytokines concentrations in the cervical mucus of pregnant women.

OBJECTIVE: Interleukin-1 alpha, interleukin-1 beta, interleukin-6 and interleukin-8 in specimens of cervical mucus were assayed to assess defense mechanisms against infection in the cervical canal during pregnancy. METHODS: Two hundred and thirty-nine pregnant women who attended the obstetrics outpatient clinic in our hospital were included as subjects. Cervical mucus was collected with a newly developed collection swab, and cytokines and granulocyte elastase in the samples of cervical mucus were quantified using enzyme immunoassay kits. The 1-factor ANOVA, the Kruskal-Wallis test, and scheffé's F were used for statistical analysis. RESULTS: Interleukin-1 alpha, interleukin-1 beta and interleukin-8 levels rose significantly as the concentrations of granulocyte elastase increased. High concentrations of interleukin-1 alpha (75.36 +/- 19.93 pg/ml), interleukin-1 beta (1005.29 +/- 238.75 pg/ml) and interleukin-8 (4822.4 +/- 633.2 pg/ml) were found in the specimens from the group of high granulocyte elastase levels. CONCLUSIONS: These cytokines might be involved in defense mechanisms against ascending infection from the vagina.

Adolescent↗
PubMed1996

Correlation between cytokine levels of amniotic fluid and histological chorioamnionitis in preterm delivery.

The aim of this study was to investigate the correlation between the cytokine levels in the amniotic fluid (AF) and the histological stage of chorioamnionitis (CAM) in premature labor. AF of 6 cases (7 samples of AF were obtained as one was a twin pregnancy) in whom CAM was diagnosed histologically, and 12 cases without CAM were included in this study. Amniotic fluid was obtained within 24 hours prior to delivery. Cytokine levels (IL-2, -4, -6, TNF-alpha, IFN-gamma) in AF were measured by an ELISA method. Levels of IL-2 and -6 in the CAM-positive group (mean +/-S.E., 52.9 +/- 83.9 pg/ml, and 20,537.9 +/- 8853.7 pg/ml, respectively) were higher than those in the CAM-negative group (i.e. undetectable, and 65.6 +/- 27.5, respectively) with a statistical significance of p < 0.05, p < 0.001, respectively. There was a positive linear relationship between IL-6 levels of AF and the placental histological inflammatory stages of Blanc in the CAM-positive group. From these results it would appear that the IL-6 level in AF is the most sensitive test in the detection of extraamniotic infection or intraamniotic infection in preterm labor with intact membranes and also indicates the severity infection.

Amniocentesis↗
PubMed1996

Two deficiency cases of disputed paternity: sibling or half-sibling?

We report briefly the results of DNA analysis in two cases of disputed paternity where the alleged father and the mother were dead and only the children were available. In one case, half-sibship was established by minisatellite typing with a multi-locus probe and microsatellite FGA typing. In the other case, Y27H39 typing revealed that the fathers of the two male children were different. DNA polymorphisms, especially at microsatellite loci, provide useful information to solve such paternity cases.

Base Sequence↗
PubMed1996

Microsatellite typing in a paternity case against a deceased man whose two brothers were available for testing.

To resolve a deficiency case of disputed paternity, we examined the mother, child, and two sibs of the deceased alleged father. In all of the 11 conventional marker systems and three microsatellite systems (ACTBP2, FGA, and D11S488) applied, the child and the paternal sibs had alleles is common. We deduced first the possible ACTBP2 genotypes of the deceased parents of the two sibs from their test results. Next, from all the combinations of the deduced genotypes, we estimated the frequencies of the possible genotypes of the alleged father and, with Bayes' theorem, calculated the probabilities of the alleged father having the genotypes compatible with paternity. Using these probabilities, we worked out the probability of the child's genotype resulting from the mating of the mother with the alleged father. Meanwhile, we calculated the probability of the child's genotype being expected of the mating of the mother with an unrelated random man. Finally, applying Bayes' theorem again, we obtained a probability of paternity of 0.992 for the ACTBP2 locus alone. Inclusion of the FGA locus in the probability calculations brought the overall probability of paternity to 0.998. The present study demonstrates that testing for hypervariable microsatellite loci greatly facilitates the solution of a deficiency case that would be difficult or time-consuming to solve by conventional marker typing.

Female↗
PubMed1996

Long-distance PCR of VNTR at the D17S74 (CMM86) locus.

We have successfully amplified D17S74 (CMM86) alleles by a long-distance polymerase chain reaction (PCR) using TaKaRa Ex Taq (a Taq DNA polymerase with a 3'-exonuclease activity) and Perfect Match Polymerase Enhancer (a special polymerase enhancer). We adopted a hot-start technique with TaqStart antibody. Because of the high guanine content (60%) in D17S74 alleles, removal of K+ from the buffers was quite effective. The use of K(+)-free buffers reduces premature chain termination in G-rich regions, thereby facilitating amplification of targets containing such sequences. The 17 alleles amplified from DNA samples of 72 unrelated Japanese subjects ranged from 1.05 to 3.5 kb, with a heterozygosity of 92%. PCR amplification of D17S74 alleles makes their detection simpler than by conventional Southern blotting, and increases the practical utility of the locus.

Alleles↗
PubMed1996

Quintuplex PCR-amplification of microsatellites.

We tested the potential usefulness of quintuplex PCR-amplification of STRs (D16S537, D8S320, FGA, D11S554 and THO1) for paternity testing, first by constructing 20 false family trios, each with a non-biological father, to estimate the actual exclusion rate, and next by a retrospective study of 20 paternity cases which had been previously tested by RFLP analysis and typing of conventional genetic markers. The five STR loci have a combined average exclusion power of 99.8%. The forensic efficiency values for four of the five STRs were based on typing of a Japanese population of 300 individuals. The observed rate of exclusion was in close agreement with the expected rate for the Japanese population. In 12 of the 20 cases, paternity was not excludable by any of the loci. The probability of paternity calculated for the alleged father was > 99% in the inclusion cases. In the remaining 8 cases, paternity was excluded by at least 2 of the STR loci. The results of STR typing agreed with the data from RFLP analysis and conventional genetic typing. Taking advantage of five STRs sharing AA in the tetranucleotide repeats (AAAG or AATG), we detected PCR products simultaneously with a digoxigenin-labeled (AAAG)6 oligonucleotide probe after sequencing-gel electrophoresis. The quintuplex PCR-amplification of STRs is a rapid and efficient method for paternity testing and individual identification.

Base Sequence↗
PubMed1996

DNA typing by triplex affinity capture (TAC) PCR.

Using a triplex affinity capture approach, we extracted PCR-ready DNA from fresh blood, buccal swabs, and bloodstains made on hard objects. We immobilized a biotinylated purine-rich oligonucleotide onto streptavidin-coated magnetic beads, and captured double-stranded DNA via triple-helix formation. We were able to amplify the MCT118, ACTBP2, FGA, D8S320, D11S488, and THO1 loci from the captured DNA. DNA typing by triplex affinity capture PCR is simple, rapid, and well-suited for paternity testing and population studies of microsatellites.

Base Sequence↗
PubMed1996

Production of monoclonal anti-thymine-dimer antibody and its use for detection of STR alleles.

To simplify the manual detection of STRs, we developed an immunoblotting method using an anti-thymine-dimer antibody. Upon ultraviolet irradiation, adjacent thymine residues in a DNA strand readily form thymine dimers which are detectable by anti-thymine-dimer antibody. We simply irradiated the supernatant of a boiled suspension of P3U1 myeloma cells with ultraviolet rays, and immunized BALB/c mice with it. By fusing their popliteal lymph node cells with the P3U1 myeloma cell line, we produced a monoclonal anti-thymine-dimer antibody. Using this antibody, we were able to detect STR alleles containing AAAG, AAGG, or AATG repeats by immunoblotting. Our method is well suited for use in parentage testing.

Alleles↗
PubMed1996

Determination of sibship by microsatellite typing in a deficiency case of disputed maternity.

In a disputed maternity case where the father and his two wives were dead and only the six children were available for testing, we analyzed 24 conventional hemogenetic markers and 11 microsatellite (STR) loci. Putting all the results together, we were able to distinguish sibship from half-sibship. Of all the systems tested, STRs were most informative. The present study demonstrates the power of STRs as a tool for solving a deficiency case.

Alleles↗
PubMed1996

STR typing of buccal swabs for paternity testing with reference to Japanese population data on the D20S85, D14S118, and D14S543 loci.

To simplify short tandem repeat (STR) typing of genomic DNA, we used buccal cells instead of blood cells. Buccal swabs taken from paternity-test trios were washed with 1 ml of distilled water by centrifugation, and the pellets were suspended in 20 microliters of distilled water. The suspensions were frozen at -80 degrees C and then thawed. Using a 1/5 volume of each suspension, we were able to type STRs at the TH01, D20S85, D14S118, and D14S543 loci by triplex PCR amplification of the first three markers and by simplex amplification of the other. In addition, we studied the polymorphisms at the D20S85, D14S118, and D14S543 loci in a Japanese population of 320 individuals. The four loci have a combined average exclusion power of 99.37%. In 24 cases of disputed paternity, the results of STR typing of the four loci agreed with those of conventional marker typing. In addition to being simple and rapid, STR typing of buccal swabs involves no painful procedure. It is therefore preferable for paternity tests on infants and small children.

Alleles↗
PubMed1996

STR typing in a deficiency case of disputed paternity.

In a case of disputed paternity where the alleged father was dead and the children and their mothers were available for testing, we analyzed 23 conventional hemogenetic markers and six short tandem repeat loci. We deduced the possible types of the alleged father and calculated the probability of paternity. Conventional phenotypings did not provide conclusive proof of paternity. In contrast, typing of the short tandem repeat loci gave a combined probability of paternity of 0.9986, establishing paternity. The present study demonstrates the power of short tandem repeat typing as a tool for solving such a deficiency case.

Alleles↗
PubMed1996

STR typing of plasma DNA in a deficiency case of disputed maternity against a patient dying in the hospital.

In this deficiency duo case, the plasma that had been left over after clinical diagnostic tests was the only blood sample available from the deceased alleged mother. Since her blood had been ABO grouped during hospitalization, we tested the plaintiff for the ABO system first. Then, to draw as much genetic information as possible from their plasma, we phenotyped 13 plasma protein systems and, three and a half years later, analyzed three short tandem repeat (STR) loci. The conventional phenotyping and STR genotyping gave probabilities of maternity of 0.974 and 0.967, respectively, bringing the total probability to 0.999. The present study highlights the utility of plasma STR typing in case of need.

ABO Blood-Group System↗
PubMed1995

Suppression of growth of renal carcinoma cells by the von Hippel-Lindau tumor suppressor gene.

Clear cell renal carcinomas are most frequently characterized by loss of function of both copies of the von Hippel-Lindau (VHL) disease gene, suggesting that the VHL gene product plays an important role in regulating renal cell proliferation. To directly assess the function of the VHL gene product, we transfected the wild-type VHL gene into two renal carcinoma cell lines that lacked normal expression of the gene. Expression of the wild-type VHL gene led to a dramatic suppression of growth in two renal carcinoma cell lines, A498 and UMRC6 in vitro, as measured by colony formation and direct cell counting. Transfection of a naturally occurring mutant VHL gene (nucleotide 713 G to A, Arg to Gln) did not lead to growth suppression of these renal carcinoma cells, nor did transfection of the wild-type VHL gene into two non-renal tumor cell lines that expressed the endogenous wild-type VHL gene. Expression constructs, which included the first ATG at nucleotide 214, were sufficient to produce the strongest growth suppression. These experiments provide direct evidence that the VHL gene product functions to suppress the growth of renal carcinoma cells and also provide a model for mapping the domains of the VHL protein important in suppressing tumor growth.

Base Sequence↗
PubMed1995

Cellular proteins that bind the von Hippel-Lindau disease gene product: mapping of binding domains and the effect of missense mutations.

The von Hippel-Lindau disease (VHL) gene is a novel tumor suppressor gene that plays a role in the pathogenesis of renal cell carcinomas and hemangioblastomas of the central nervous system. To begin an evaluation of the biological functions of the VHL gene product (pVHL), we prepared bacterial fusion protein between glutathione S-transferase and wild-type or mutant pVHLs. The fusion proteins were used to identify cellular proteins that bind to pVHL in vitro. Monkey kidney cells transfected with wild-type or mutant VHL cDNAs were used to identify cellular proteins that bind to pVHL in vivo. Wild-type pVHL consistently bound two cellular proteins with apparent molecular masses of 10 and 14 kilodaltons that were designated p10 and p14, respectively. Mapping studies with a panel of VHL deletion mutant proteins demonstrated that p10 and p14 bound to a 32-amino acid peptide located in the carboxy terminal portion of pVHL. Missense mutation located within this 32-amino acid peptide abrogated the ability of the VHL protein to bind p10 and p14. Of 67 VHL families with identified germline mutations, 42 families had mutations predicted to affect the p10/p14-binding region. Maintenance of the integrity of the p10/p14-binding region appears to be essential for cellular growth regulation by pVHL.

Animals↗